FRET-based homogeneous immunodetection method and detection composition

A composition and bismuth-based technology, applied in the field of analytical chemistry, can solve the problems of high background fluorescence and low sensitivity, and achieve the effects of high detection sensitivity, little background fluorescence interference and simple method

Pending Publication Date: 2022-03-04
CHANGCHUN INST OF OPTICS FINE MECHANICS & PHYSICS CHINESE ACAD OF SCI
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In view of this, the object of the present invention is to provide a FRET-based homogeneous immunoassay method and detection composition to solve the technical problems of low sensitivity and high background fluorescence existing in the existing FRET-based homogeneous immunoassay method

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  • FRET-based homogeneous immunodetection method and detection composition
  • FRET-based homogeneous immunodetection method and detection composition
  • FRET-based homogeneous immunodetection method and detection composition

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preparation example Construction

[0053] In the present invention, the bismuth-based material is a bismuth-based two-dimensional nano-sheet material or a bismuth-based two-dimensional nano-sheet material doped with metal ions. This material is a prior art, and can be obtained by methods familiar to those skilled in the art such as commercial purchase or laboratory synthesis. For example, the commonly used bismuth-based two-dimensional nanosheet material is Bi 2 S 3 、 Bi 2 Se 3 or Bi 2 Te 3 , the present invention preferably bismuth-based two-dimensional nanosheet material Bi 2 Se 3 or Bi 2 Te 3 . The doped metal ions can be rare earth ions, transition metal ions, etc., and the doping ratio is not limited. The present invention provides a method for preparing a bismuth-based material labeled with an antigen, but is not limited thereto. Specifically, the bismuth-based material is stirred and mixed with a solution containing an antigen for more than 12 hours, then centrifuged, and the obtained precipita...

Embodiment 1

[0070] 1) Bismuth-based materials (Bi 2 Se 3 preparation of nanomaterials)

[0071] 0.372mmolBi(NO 3 ) 3 ·.3H 2 O, 0.560mmol NaSeO 3 And put 0.4g of PVP into a three-necked bottle with a capacity of 50mL, and add 24mL of ethylene glycol. Then put the three-necked bottle into a magnetic stirring heater, and at the same time pass through nitrogen for protection, heat to 60 ° C, and maintain at this temperature for 30 min, after the solid is completely dissolved, a clear and transparent solution is obtained. Then heat the clear and transparent solution to 180°C, inject a mixed solution of 0.8mL hydrazine hydrate and 8mL ethylene glycol at this temperature, and react for 30 minutes. The reaction is carried out under nitrogen protection, and cooling water is used to reflux. to room temperature, purified once by centrifugation with acetone to obtain Bi 2 Se 3 nanomaterials, the resulting Bi 2 Se 3 Nanomaterials were dispersed into 16mL deionized water and set aside.

[00...

Embodiment 2

[0086] Step 1), 3), 4), 5) are the same as in Example 1, and 2 in Example 1) is replaced by:

[0087] 1mmol of rare earth chlorides (including 0.745mmol YCl 3 ·6H 2 O, 0.25 mmol YbCl 3 ·6H 2 O and 0.05 mmol TmCl 3 ·6H 2 O) put into capacity and be in the three-necked bottle of 50mL, add the oleic acid of the octadecene of 15mL and 6mL. Then put the three-necked bottle into a magnetic stirring heater, and at the same time pass through argon gas for protection, and heat it to 160°C until the chloride is completely dissolved to obtain a slightly light yellow, clear and transparent solution. Then cool to room temperature, dissolve 4mmol of NH 4 F. Add 2.5 mmol of LiOH in 10 mL of methanol solution dropwise to the above-mentioned three-necked flask, then heat to 75°C and keep for 30 minutes to completely remove methanol in the solution, then heat to 300°C and react for 1.5h. During the process, the argon gas was maintained for protection, and cooling water was used to reflux...

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Abstract

The invention relates to an FRET (Fluorescence Resonance Energy Transfer)-based homogeneous immunodetection method and a detection composition, and belongs to the technical field of analytical chemistry. The technical problems that an existing FRET-based homogeneous immunodetection method is low in sensitivity, high in background fluorescence and the like are solved. The detection composition comprises a bismuth-based material marked with an antigen and a rare earth doped luminescent nanocrystal marked with an antibody, or the bismuth-based material marked with the antibody and the rare earth doped luminous nanocrystal marked with the antigen are included; wherein the antigen and the antibody are antigen-antibody pairs capable of carrying out specific binding reaction; the bismuth-based material is a bismuth-based two-dimensional nanosheet material or a metal ion doped bismuth-based two-dimensional nanosheet material. The invention also provides a method for antibody or antigen detection by using the detection composition. The FRET-based homogeneous immunodetection method has the advantages of small background fluorescence interference, high detection sensitivity, low cost, simplicity, convenience in operation and the like.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry, and in particular relates to a FRET-based homogeneous immunoassay method and a detection composition, which can be used for antigen detection and antibody detection. Background technique [0002] The homogeneous immunoassay method based on fluorescence resonance energy transfer (FRET) is a research and application hotspot in recent years. FRET refers to the non-radiative energy transfer process between a donor and an acceptor within a certain distance (generally <10nm). This spatial confinement effect endows FRET homogeneous detection technology with many advantages, such as no need for separation and purification of the analyte, simple operation method, fast detection speed, good selectivity, wide application range, and simultaneous multi-component detection, etc. . It can be seen that the selection of donors and acceptors is very important to realize the above-mentioned rapid ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/533
CPCG01N33/582G01N33/587G01N33/533
Inventor 常钰磊解小雨高丹丹曲林琳孔祥贵
Owner CHANGCHUN INST OF OPTICS FINE MECHANICS & PHYSICS CHINESE ACAD OF SCI
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